microRNA Regulation of Gamma-herpesvirus Latency and Reactivation

项目摘要 EB病毒（EBV）持续感染全世界超过90%的成年人。在患有先天性或后天性 免疫缺陷，如接受器官移植或艾滋病毒/艾滋病患者，终身持续 感染可导致多种癌症。像所有疱疹病毒一样，EBV具有潜伏性和裂解性复制 阶段;在这些程序之间切换的能力是病毒长期持续存在的关键。周期性的，潜伏的 EBV重新激活产生感染性病毒，这被认为是增加感染细胞的池，并维持 持续感染调节潜伏期和向裂解性复制转变的分子机制并不完全 明白microRNAs（miRNAs）是一类非编码的小分子调控RNA，它调控着许多生物学过程， 包括细胞状态转换和免疫反应的发展过程。EBV编码 进化上保守的病毒miRNAs在病毒生命周期的各个阶段都有活性， 高度限制性的，可以干扰宿主信号传导途径和抗病毒防御机制， 多个水平，这使我们假设病毒miRNAs代表了为什么宿主免疫反应的重要组成部分， 无法清除持续感染。尽管我们做出了巨大的努力来促进对EBV miRNA功能的理解， 关于它们在体内对感染的作用知之甚少。根据初步的发现，我们假设， EBV miRNAs作为毒力因子，协调潜伏期和再激活方面，促进病毒的传播。 在宿主中的持久性。在这个项目中，我们将研究调控非编码RNA的相互作用， 特别是病毒和细胞miRNAs如何在EBV感染过程中协同塑造动态细胞状态， 确定影响病毒再活化的关键决策的miRNAs。我们的目标是阐明关键的 miRNA和miRNA介导的机制（靶点和途径）调节EB病毒潜伏性向裂解性转变。到 为了验证我们关于病毒miRNAs促进体内持久性的假设，我们将利用恒河猴淋巴隐病毒 在自然灵长类宿主中检查病毒miRNA表型的模型。本文提出的研究具有实用性， 开发基于RNA的抗病毒性疾病的治疗策略，并对推进基因治疗具有重要意义。 疱疹病毒疫苗开发。

PROJECT SUMMARY Epstein-Barr virus (EBV) persistently infects >90% of adults worldwide. In individuals with congenital or acquired immune deficiencies, such as patients undergoing organ transplant or living with HIV/AIDS, life-long persistent infection can lead to a variety of cancers. Like all herpesviruses, EBV has both a latent and a lytic replication phase; the ability to switch between these programs is key to long-term viral persistence. Periodically, latent EBV reactivates to produce infectious virus, which is thought to increase the pool of infected cells and sustain persistent infection. Molecular mechanisms regulating latency and the switch to lytic replication are not entirely understood. MicroRNAs (miRNAs) are small, non-coding regulatory RNAs that govern many biological processes including cell state transitions and the development of immune responses. EBV encodes evolutionarily conserved viral miRNAs that are active during stages of the viral life cycle when viral proteins are highly restricted and that can interfere with host signaling pathways and anti-viral defense mechanisms on multiple levels, leading us postulate that viral miRNAs represent an important part of why host immune responses fail to clear persistent infection. Despite significant efforts advancing our understanding of EBV miRNA functions, little is known about their contributions to infection in vivo. Based on preliminary findings, we hypothesize that EBV miRNAs act as virulence factors and coordinate aspects of latency and reactivation that facilitate virus persistence within a host. In this project, we will examine regulatory non-coding RNA interactions, asking specifically how viral and cellular miRNAs cooperatively shape dynamic cell states during EBV infection, and determine the miRNAs influencing critical decisions that impact virus reactivation. We aim to elucidate the critical miRNAs and miRNA-mediated mechanisms (targets and pathways) regulating the EBV latent to lytic switch. To test our hypothesis that viral miRNAs facilitate persistence in vivo, we will leverage the rhesus lymphocryptovirus model to examine viral miRNA phenotypes in a natural primate host. Studies proposed herein have utility in developing RNA-based therapeutic strategies against viral disease and important implications for advancing g- herpesvirus vaccine development.